Universal Journal of Pharmacy Take Research to New Heights
|
|
- William Austin
- 8 years ago
- Views:
Transcription
1 Universal Journal of Pharmacy Take Research to ew Heights Research Article ISS X SYTHESIS, ATI-IFLAMMATRY, ATIMICRBIAL AD ATHELMITIC EVALUATI F AXIASTATI-1 *Corresponding author: Dr. (Mrs.) M. Himaja Pharmaceutical Chemistry Division, School of Advanced Sciences, VIT University, Vellore , Tamil adu, India Mobile: , Malipeddi Himaja * * Pharmaceutical Chemistry Division, School of Advanced Sciences, VIT University, Vellore , Tamil adu, India Received ; Revised ; Accepted ABSTRACT Axinastatin 1, a cyclic heptapeptide, was synthesized by solution phase peptide synthesis using -(Benzotriazol-1-yl)-,,, -tetramethyluronium tetrafluoroborate (TBTU) as the coupling agent and triethylamine as the base. The structure of the compound was confirmed by FTIR, 1 H MR, FABMASS and elemental analysis. The synthesized cyclic peptide was evaluated in vitro anti-inflammatory activity by membrane stabilizing method and antimicrobial activity against four strains of bacteria and two strains of fungi. It was also evaluated for anthelmintic activity against earthworms. The compound showed moderate anti-inflammatory activity as compared to the standard drug, ibuprofen. It possessed potent antibacterial activity only against Gram positive bacteria and moderate anthelmintic activity as compared to the standard drug, mebendazole. Keywords: Axinastatin 1, Solution phase peptide synthesis, Anti-inflammatory activity, Antimicrobial activity, Anthelmintic activity. ITRDUCTI Peptides belongs to one of the most important classes of organic compounds possessing many biological activities like antioxidant, anthelmintic, antitubercular, anti-inflammatory 1-3, antifungal and antibacterial activities 4,5. The objective in peptide synthesis is to design peptide ligands with specific physical, chemical and biological activities. Peptides ligands generally act by interaction with acceptor or receptor molecules (enzymes, hormones, neurotransmitters, etc.) 6,7. Axinastatin 1 is a cyclic peptide, possessing relatively simple chemical structure. It is a cyclic heptapeptide, produced by the Western Pacific marine sponge Axinella species. It was first isolated by George Pettit et al. in the year The sponge was collected in Palau and extracted with a mixture of methylene chloride and 2-propanol (1:1). Axinastatin 1 was crystallized from methylene chloride. Axinastatin 1 exhibited cytotoxicity against the PS leukemia cell line. The structure of Axinastatin 1 consists of three L-Valine, one L- Phenylalanine, two L-Proline and one L-Asparagine units (Figure 1). In order to carry out the synthesis, the cyclic heptapeptide was disconnected into three dipeptide units and a single amino acid unit. The dipeptides were prepared from the respective protected amino acids. The amino group was protected with tertiary Butyloxycarbonyl (Boc-) group and the carboxyl group was protected by converting it into methyl ester. The Boc-amino acids were coupled with the amino acid methyl ester hydrochlorides by using -(Benzotriazol-1-yl)-,,, -tetramethyluronium tetrafluoroborate (TBTU) as the coupling agent and triethylamine (TEA) as the base. Universal Journal of Pharmacy, 04(05), September-ctober H 3 C C H 3 H 2 CH 3 H CH H 3 C 3 CH 3 Figure 1: Structure of Axinastatin 1
2 MATERIALS AD METHDS All the reactions requiring anhydrous conditions were conducted in flame dried apparatus. All the reactions were magnetically stirred unless otherwise stated. rganic extracts were dried over anhydrous sodium sulphate. Melting points were determined by capillary method and were uncorrected. Amino acids, di-tertbutylpyrocarbonate, trifluoroacetic acid and triethylamine were obtained from Spectrochem Ltd. Mumbai. TBTU, DCM, diethyl ether, methanol and chloroform was obtained from AVRA. IR spectra were recorded on Perkin-Elmer IR spectrometer using a thin film supported on KBr pellets for solids and chloroform as a solvent for semisolids. The values are reported as ν max (cm 1 ). 1 H MR spectra were recorded on Bruker JEL (400MHz) MR spectrometer. The spectra were obtained in CDCl 3 and the chemical shift values are reported as values in ppm relative to TMS (δ = 0) as internal standard. FABMASS spectra were recorded on a Joel Sx 102/DA-6000 mass spectrometer using xenon as the carrier gas. The spectra were recorded at room temperature, m-nitrobenzyl alcohol was used as the matrix. The protection of amino group, carboxyl group and side chain and their deprotection were done by standard procedures 9,10. Preparation of Dipeptides 10 mmol of amino acid methyl ester hydrochloride was dissolved in 20 ml of dichloromethane (DCM). To this, triethylamine (TEA) (4 ml, 28.7 mmol) was added at 0 C and the reaction mixture was stirred for 15 mins. Boc-amino acid (10 mmol) in DCM (20 ml) and TBTU (10 mmol) were added with stirring. After 1 h, the reaction mixture was filtered and the residue was washed with DCM (30 ml). The filtrate was washed with 5% ahc 3 (20 ml), 5% HCl (20 ml) and distilled H 2 (20 ml). The organic layer was dried over anhydrous a 2 S 4, filtered and evaporated in a vacuum. The residue was purified by recrystallization from DCM. Boc-Pro-Phe-Me, Boc- Val-Val-Me and Boc-Pro-Val-Me was prepared in this manner. Protection of the Side chain function The carboxamide side function of Boc-Asparagine was protected by introducing the benzhydryl group (bzh) by Bodanszky 10 method (Scheme 1). Preparation of Axinastatin 1 The ester group of Boc-Val-Val-Me was removed with LiH and the Boc- group of Boc-Pro-Val-Me 2 was removed by trifluoroacetic acid (TFA). The deprotected dipeptides were coupled using TBTU and TEA to get the tetrapeptide Boc-Val-Val-Pro- Val-Me. The Boc- group of the tetrapeptide and the ester group of the dipeptide Boc-Pro-Phe-Me were removed coupled to give the hexapeptide Boc-Pro-Phe-Val-Val-Pro-Val-Me. After the removal of the Boc- group of the hexapeptide unit, it was coupled with Boc-Asn(bzh)-H to obtain the heptapeptide Boc-Asn(bzh)-Pro-Phe-Val-Val-Pro- Val-Me. The ester group of the heptapeptide was removed and p-nitrophenyl group (pnp) was introduced. The Boc- and the bzh- groups were removed by TFA and the linear fragment was cyclized by adding pyridine and keeping the whole contents at 0 o C for ten days to get the Axinastatin 1 (Scheme 2). In vitro Anti-inflammatory Activity In vitro Anti-inflammatory Activity of Axinastatin 1 was carried out by the below mentioned procedures 11. Preparation of erythrocyte suspension 10 ml of blood was collected and centrifuged at 3000 rpm for 10 mins in a centrifuge. Plasma was discarded and the pellet was washed with 0.9% saline. The volume of the blood was reconstituted as 20% v/v suspension with 10 mm sodium phosphate buffer solution (ph 7.4). Heat induced haemolysis The membrane stabilizing activity of Axinastatin 1 was measured by heat induced haemolysis of human blood. The reaction mixture contained solutions of Axinastatin 1 ( µg/ml) in different tubes. 20% RBCs suspensions (1 ml) was added to all the tubes. Instead of the compound only 10 mm sodium phosphate buffer solution was added to the control tube. The tubes containing the mixture were incubated at 56 o C for 30 minutes in a water bath. The reaction mixture was centrifuged at 3000 rpm for 10 minutes and the absorbance of the supernatants was measured at 560 nm using Hitachi U-2910 Spectrophotometer. Ibuprofen ( µg/ml) was used as the reference standard. Membrane stabilizing activity (in %) was calculated by the following formula: Inhibition (%) = 100 Where, A t = Absorbance of test sample and A c = Absorbance of control. (bzh) Boc-H CH 3 CH, benzhydrol conc. H 2 S 4, RT, 12 h Boc-H H H Boc-Asn(bzh) Scheme 1: Protection of side chain of Asparagine Universal Journal of Pharmacy, 04(05), September-ctober
3 Boc-Pro + Phe-Me Boc-Val + Val-Me Boc-Pro + Val-Me a, b Boc-Pro-Phe + a, b Boc-Val-Val + Val-Val-Pro-Val-Me Pro-Val-Me Pro-Phe-Val-Val-Pro-Val-Me a Boc-Asn(bzh) Boc-Asn(bzh)-Pro-Phe-Val-Val-Pro-Val-Me a = TBTU, TEA, DCM, RT, 1 h b = LiH, THF:H 2 (1:1), RT, 1 h b, d, c, e c = TFA, CHCl 3, RT, 1 h d = p-nitrophenol, CHCl 3, RT, 12 h Axinastatin 1 e = Pyridine, CHCl 3, 10 days, 0 o C Scheme 2: Synthetic scheme of Axinastatin 1 Antimicrobial Activity For antimicrobial activity study four strains of bacteria and two strain of fungi were used. The bacterial isolates included two Gram negative bacteria (Pseudomonas aeruginosa and Escherichia coli) and two Gram positive bacteria (Staphylococcus aureus and Bacillus subtilis). The fungal cultures were Candida albicans and Aspergillus niger. The antimicrobial activity of Axinastatin 1 was tested at 50 µg/ml, using agar well diffusion assay method 12. The bacterial test organisms were grown in nutrient broth at 37 o C for 24 h and then spread on Muller Hinton Agar (MHA) plates. For the fungal cultures, spore suspensions was grown in Sabor Dextrose Broth (SDB) at 25 o C for 48 h and then spread on Potato Dextrose Agar (PDA) plates. Dimethyl formamide was used as negative control and disks containing penicillin (10µg/disk), streptomycin (10µg/disk) and griseofulvin (25µg/disk) as positive controls. All the tests were performed in triplicates. The antimicrobial activity was evaluated based on the diameter of zone of inhibition. Anthelmintic Activity Anthelmintic activity study of Axinastatin 1 was carried out against earthworms (Eudrilus eugeniae 13 ). Suspension of Axinastatin 1 was prepared by triturating it with 15% Tween 80 and distilled water and the mixture was stirred for 30 mins using a magnetic stirrer. The suspension was diluted to contain 0.1% and 0.2% w/v in 20 ml of the test sample. The standard drug, mebendazole was also prepared in a similar way. Earthworms were placed in three petri dishes containing 20 ml of each sample, control (20 ml suspension of distilled water and 15% Tween 80) and standard drug, respectively at room temperature. The time needed for the paralysis and death of the earthworms were noted. The death time was determined by placing the earthworms in warm water at 50 o C, which stimulated the movement if the earthworms were alive. RESULTS AD DISCUSSI Physical Data and Spectral Analysis Axinastatin 1: Yield 69%; pale white solid; m.p o C; IR spectrum (ν/cm -1 ): 3300 (br. s), 2930 (s), 2965 (s), 1705 (s), 1690 (s), 1670 (br. s), 1645 (br. s), 1620 (s), 1520 (s), 1470 (s), 1450 (s), 1280 (s), 1165 (s), 1110 (s), 1040 (s), 895 (s) cm -1 ; 1 H MR spectrum (δ, ppm): 8.5(3H, br. s), 7.5 (4H, br. s),, (5H, m), (1H, m), (2H, m), (4H, m), (6H, m), (10H, m), (3H, m), 1.0 (18H, d, J = 6.0 Hz); FABMASS: m/z (M + H) + = 753.4; Elemental Analysis: M. F. = C 38 H , M. W. = 752, Found (Cal) %C: (60.64), %H: 7.46 (7.45), %: (14.89). Anti-inflammatory Activity Anti-inflammatory activity of Axinastatin 1 are reported in Table 1. From the obtained results it was concluded that Axinastatin 1 has a significant membrane stabilizing activity which was comparable to the standard ibuprofen. The maximum inhibition of 71.3±0.69% was observed with 200 µg/ml of Axinastatin 3. Whereas, diclofenac sodium, showed the maximum inhibition of 89.4±0.63% at 200 µg/ml. Universal Journal of Pharmacy, 04(05), September-ctober
4 Table 1: In vitro anti-inflammatory activity of Axinastatin 1 Concentration (µg/ml) Standard (ibuprofen) Axinastatin ± ± ± ± ± ±0.69 Values are expressed as mean ± S. E of the three replicates. Antimicrobial Activity Evaluation of antibacterial and antifungal activities was done for Axinastatin 1 (Table 1) and it was found to possess antibacterial activity against Gram positive (B. subtilis and S. aureus) bacteria. However, it did not show antibacterial activity against Gram negative (P. aeruginosa and E. coli) bacteria and antifungal activity against C. albicans and A. niger. Table 2: Antimicrobial activity of Axinastatin 1 Diameter of zone of inhibition (in mm). P. aer E. coli B. sub S. aur C. alb A. niger Sl. o. Compound 1 Axinastatin ± ± Penicillin 11.2± ± ± ± Streptomycin 16.2± ± ±1 12.2± Griseofulvin ± ±1 Values are expressed as mean ± S. E of the three replicates. Zone of inhibition not include the diameter of the well (7 mm) Anthelmintic activity The synthesized cyclic peptide, Axinastatin 1 have shown moderate anthelmintic activity as compared to CCLUSI the standard drug, mebendazole. Table 2 illustrates the anthelmintic activity of sample and standards. Table 3: Anthelmintic activity of Axinastatin 1 Sl. o. Compound Conc. of the Mean paralysing time Mean death time compound (mg) (mins) ± S.E (mins) ± S.E 1 Control Mebendazole ± ± ± ± Axinastatin ± ± ± ± 1.10 Axinastatin 1 can be easily synthesized under normal laboratory conditions. The synthesis is cost effective and time saving. The structure of the cyclic peptide was confirmed by IR, 1 H MR, FABMASS and elemental analysis. The compound was screened for in vitro antiinflammatory activity and it showed potent antiinflammatory activity as compared to the standard, ibuprofen. Axinastatin 1 was screened for antimicrobial activity against four strains of bacteria and two strains of fungi and it showed potent antibacterial activity only against the Gram positive bacteria. The compound was also screened for anthelmintic activity and it showed moderate activity as compared to the standard drug, mebendazole. ACKWLEDGEMETS We are grateful to the VIT-SIF Lab, School of Advanced Sciences, VIT University, Vellore, for the support of this research. REFERECES 1. Das M. and Himaja M., Synthesis, antioxidant and anthelmintic activity of the linear tetrapeptide L-(Leu-Pro-Gly)-D-Ala (LPGA), Int. J. Pharm. Pharm. Sci., 2013, 5(Suppl 3), Das M, Agarwal DS, Sharma SK, Das P and Rout PK., Synthesis, docking studies and free radical scavenging activity of the linear tetrapeptide VFPF, Int. Res. J. Pharm., 2012; 3(6): Himaja M, Sreekanth K., Munirajsekhar D, Ramana MV. and Mukesh S, Computer- aided design, synthesis and antioxidant activity of linear tetrapeptide D-Phe-L-(Ala-Tyr-Val), J. Pharm. Res. 2011; 4(8): Tadahiro T, Tayuka K, oriko S, Yukio and Machiko M, Synthesis of triglycosyl tetrapeptides and a hexaglycosyl tetrapeptide, Carbohydr. Res., 1996, 283, Daniele B, Andrea B, Matteo C, Gianni P and Sergio S, Synthesis and conformational preferences of unnatural tetrapeptides Universal Journal of Pharmacy, 04(05), September-ctober
5 containing L-valine units, Tetrahed: Asymm., 2006; 17: Victor JH, Fahad A and Wieslaw K., Emerging approaches in the molecular design of receptor-selective peptide ligand, conformational, topographical and dynamic considerations, Biochem. J., 1990; 268: David S, Lenka Z and Milos B, High performance liquid chromatography and nuclear magnetic resonance study of linear tetrapeptides and octapeptides containing -methylated amino acid residues, J. Chromatogr, A, 2007; 1160: Petit GR, Herald CL, Boyd MR, Leet JE, Dufresne C, Dennis LD, Schmidt JM, Cerny RL, Hooper JA and Rutzler KC, Isolation and structure of the cell growth inhibitory constituents from the western Pacific marine sponge Axinella sp., J. Med. Chem., 1991; 34: Dahiya R and Gautam H., Total synthesis and antimicrobial activity of a natural cycloheptapaptide of marine origin, Marine Drugs, 2010, 8, Bodanszky M and Bodanszky A., Practice of peptide synthesis, ew York: Springer-Verlag, 1984, p Shinde UA, Phadke AS, ari AM, Mungantiwar AA, Dikshit VJ and Saraf M., Membrane stabilization activity - A possible mechanism of action for the anti-inflammatory activity of Cedrusdeodara wood oil, Fitoterapia, 1999; 70: Rahman MM, Mosaddik MA, Wahed MII and Haque ME., Antimicrobial activity and cytotoxicity of Trapa Bispinosa, Fitoterapia, 2010; 71: Garg LC and Atal CK, Evaluation of Anthelmintic activity, Ind. J. Pharmacol., 1969; 32: 104. Source of support: il, Conflict of interest: one Declared Universal Journal of Pharmacy, 04(05), September-ctober
Unique Research Journal of Chemistry DOCKING, SYNTHESIS AND BIOLOGICAL EVALUATION OF A LINEAR TETRAPEPTIDE L-GLFP
ISS 2348-232X Unique Research Journal of Chemistry Available online: www.ujconline.net Research Article DCKIG, SYTHESIS AD BILGICAL EVALUATI F A LIEAR TETRAPEPTIDE L-GLFP Kavitha P 1*, Mahima M 1, Saravanan
More informationSynthesis and Antimicrobial Activity of 7-Amino cephalosporanic Acid Derivatives of Amino Acids and Peptides
Asian Journal of Chemistry Vol. 22, No. 4 (2010), 2914-2918 Synthesis and Antimicrobial Activity of 7-Amino cephalosporanic Acid Derivatives of Amino Acids and Peptides M. HIMAJA*, SIDDHARTH DESAI, A.
More informationSynthesis and Biological screening of Proline rich Cyclic Heptapeptide
International Journal of PharmTech Research CDE (USA): IJPRIF ISS : 0974-4304 Vol.4, o.1, pp 194-200, Jan-Mar 2012 Synthesis and Biological screening of Proline rich Cyclic eptapeptide Sachin Chaudhary
More informationFirst total synthesis and biological screening of cyclic tetrapeptides from onychocola sclerotica
Available online at wwwderpharmachemicacom Scholars Research Library Der Pharma Chemica, 15, 7(11):251-257 (http://derpharmachemicacom/archivehtml) ISS 975-413X CDE (USA): PCAX First total synthesis and
More informationSupporting Information
Copyright WILEY VCH Verlag GmbH & Co. KGaA, 69469 Weinheim, Germany, 2012. Supporting Information for Adv. Funct. Mater., DOI: 10.1002/adfm.201102486 Colorimetric Detection of Warfare Gases by Polydiacetylenes
More information4026 Synthesis of 2-chloro-2-methylpropane (tert-butyl chloride) from tert-butanol
4026 Synthesis of 2-chloro-2-methylpropane (tert-butyl chloride) from tert-butanol OH + HCl Cl + H 2 O C 4 H 10 O C 4 H 9 Cl (74.1) (36.5) (92.6) Classification Reaction types and substance classes nucleophilic
More informationAcid-Base Extraction.
Acid-Base Extraction. Extraction involves dissolving a compound or compounds either (1) from a solid into a solvent or (2) from a solution into another solvent. A familiar example of the first case is
More informationExperimental procedures. Solid phase peptide synthesis (SPPS)
Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry This journal is The Royal Society of Chemistry 214 Experimental procedures Solid phase peptide synthesis (SPPS) Solid phase
More informationA Ratiometric NMR ph Sensing Strategy Based on Slow- Proton-Exchange (SPE) Mechanism
Electronic Supplementary Material (ESI) for Chemical Science. This journal is The Royal Society of Chemistry 2015 A Ratiometric NMR ph Sensing Strategy Based on Slow- Proton-Exchange (SPE) Mechanism Loïse
More informationExperiment 3: Extraction: Separation of an Acidic, a Basic and a Neutral Substance
1 Experiment 3: Extraction: Separation of an Acidic, a Basic and a Neutral Substance Read pp 142-155, 161-162, Chapter 10 and pp 163-173, Chapter 11, in LTOC. View the videos: 4.2 Extraction (Macroscale);
More informationand its application in the synthesis of Nilotinib intermediate
Electronic upplementary Material (EI) for RC Advances. This journal is The Royal ociety of Chemistry 204 upporting Information An efficient D-glucosamine-based copper catalyst for C-X couplings and its
More informationOverview'of'Solid-Phase'Peptide'Synthesis'(SPPS)'and'Secondary'Structure'Determination'by'FTIR'
verviewofsolid-phasepeptidesynthesis(spps)andsecondarystructuredeterminationbyftir Introduction Proteinsareubiquitousinlivingorganismsandcells,andcanserveavarietyoffunctions.Proteinscanactas enzymes,hormones,antibiotics,receptors,orserveasstructuralsupportsintissuessuchasmuscle,hair,and
More informationMicrowave irradiated high-speed solution synthesis of peptide acids employing Fmoc-amino acid pentafluorophenyl esters as coupling agents
Indian Journal of Chemistry Vol. 44B, ovember 2005, pp. 2328-2332 Microwave irradiated high-speed solution synthesis of peptide acids employing Fmoc-amino acid pentafluorophenyl esters as coupling agents
More informationA prochelator with a modular masking group featuring hydrogen peroxide activation with concurrent fluorescent reporting
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2014 Supporting Information A prochelator with a modular masking group featuring hydrogen peroxide activation
More informationPeptide Synthesis Zheng Miao* and Zhen Cheng
Peptide Synthesis Zheng Miao* and Zhen Cheng 1 Department of Radiology, Molecular Imaging Program at Stanford, Stanford University School of Medicine, Stanford, USA *For correspondence: zmiao@stanford.edu
More informationNMR Chemical Shifts of Common Laboratory Solvents as Trace Impurities
7512 J. Org. Chem. 1997, 62, 7512-7515 NMR Chemical Shifts of Common Laboratory Solvents as Trace Impurities Hugo E. Gottlieb,* Vadim Kotlyar, and Abraham Nudelman* Department of Chemistry, Bar-Ilan University,
More informationAlkoxycarbonylation of Ethylene with Cellulose in Ionic Liquids
Supplementary data Alkoxycarbonylation of Ethylene with Cellulose in Ionic Liquids Anna sichow and Stefan Mecking* Chair of Chemical Materials Science, Dept. of Chemistry, University of Konstanz, 78464
More information1) Technical informations. - a) How does it work? - b) Purification - c) Quality Control. 2) Standard synthesis
1) Technical informations - a) How does it work? - b) Purification - c) Quality Control 2) Standard synthesis - a) Standard peptides - b) Modified peptides - c) Shipment and Delivery Time - d) How to order?
More informationSmall μmol Scale Synthesis of a Labeled Antimicrobial Peptide using Biotage
Application ote A098 Small μmol Scale Synthesis of a Labeled Antimicrobial Peptide Page 1 Small μmol Scale Synthesis of a Labeled Antimicrobial Peptide using Biotage Initiator+ Alstra Introduction Labeled
More informationNovel Method for Solid Phase Peptide Synthesis Using Microwave Energy
Novel Method for Solid Phase Peptide Synthesis Using Microwave Energy Jonathan M. Collins, Michael J. Collins, Rebecca C. Steorts CEM Corporation, Matthews, NC 28106-0200, U.S.A. Presented at American
More informationSupplementary Information for
Electronic Supplementary Material (ESI) for Polymer Chemistry. This journal is The Royal Society of Chemistry 2015 Supplementary Information for Doubly Responsive Polymersomes towards Monosaccharides and
More informationEUDRAGIT E 100, EUDRAGIT E PO and
Technical Information EUDRAGIT E 100, and Specification and Test Methods Ph. Eur. USP/NF JPE Basic Butylated Methacrylate Copolymer Amino Methacrylate Copolymer - NF Aminoalkyl Methacrylate Copolymer E
More informationUTILIZATION of PLASMA ACTIVATED WATER in Biotechnology, Pharmacology and Medicine. JSC TECHNOSYSTEM-ECO Moscow, Russia April, 2009
UTILIZATION of PLASMA ACTIVATED WATER in Biotechnology, Pharmacology and Medicine JSC TECHNOSYSTEM-ECO Moscow, Russia April, 2009 METHOD of WATER ACTIVATION with PLASMA of GAS DISCHARGE ANODE VACUUM WATER
More information1. COUPLING REAGENTS : Structure and acronyms
Coupling Reagents 1. COUPLING REAGENTS : Structure and acronyms... 2 2. CARBODIIMIDE... 3 1.a. N,N -Dicyclohexylcarbodimide (DCC)... 3 DCC/HOBt coupling experimental procedure:... 4 1.b. N-(3-Dimethylaminopropyl)-N
More informationThe D-glucosamine-derived pyridyl-triazole@palladium recoverable catalyst for Mizoroki-Heck reactions under solvent-free conditions
Electronic Supplementary Material (ESI) for Green Chemistry. This journal is The Royal Society of Chemistry 204 Supporting Information The D-glucosamine-derived pyridyl-triazole@palladium recoverable catalyst
More informationThe Synthesis of trans-dichlorobis(ethylenediamine)cobalt(iii) Chloride
CHEM 122L General Chemistry Laboratory Revision 2.0 The Synthesis of trans-dichlorobis(ethylenediamine)cobalt(iii) Chloride To learn about Coordination Compounds and Complex Ions. To learn about Isomerism.
More informationCHEM 2423 Recrystallization of Benzoic Acid EXPERIMENT 4 - Purification - Recrystallization of Benzoic acid
EXPERIMENT 4 - Purification - Recrystallization of Benzoic acid Purpose: a) To purify samples of organic compounds that are solids at room temperature b) To dissociate the impure sample in the minimum
More informationEffects of Antibiotics on Bacterial Growth and Protein Synthesis: Student Laboratory Manual
Effects of Antibiotics on Bacterial Growth and Protein Synthesis: Student Laboratory Manual I. Purpose...1 II. Introduction...1 III. Inhibition of Bacterial Growth Protocol...2 IV. Inhibition of in vitro
More informationlung cancer targeted photodynamic therapy and imaging
99m Tc-Hematoporphyrin linked albumin nanoparticles for lung cancer targeted photodynamic therapy and imaging Su-Geun Yang, Ji-Eun Chang, Byungchul Shin, Sanghyun Park, Kun Na and Chang-Koo Shim* *Corresponding
More informationChemistry 321, Experiment 8: Quantitation of caffeine from a beverage using gas chromatography
Chemistry 321, Experiment 8: Quantitation of caffeine from a beverage using gas chromatography INTRODUCTION The analysis of soft drinks for caffeine was able to be performed using UV-Vis. The complex sample
More informationSyllabus CHM 2202 Organic Chemistry Laboratory II Spring 2011
Villanova University Department of Chemistry Syllabus CHM 2202 Organic Chemistry Laboratory II Spring 2011 Text: C.E. Bell, D.F. Taber and A.K. Clark, Organic Chemistry Laboratory with Qualitative Analysis,
More informationISOLATION OF CAFFEINE FROM TEA
ISLATIN F CAFFEINE FRM TEA Introduction In this experiment, caffeine is isolated from tealeaves. The chief problem with the isolation is that caffeine does not exist alone in the tealeaves, but other natural
More informationEvaluation of Microbial Growth and Survival on Construction materials treated with Anabec NewBuild 30
Evaluation of Microbial Growth and Survival on Construction materials treated with Anabec NewBuild 30 Absar Alum, Ph.D. Department of Civil and Environmental Engineering Arizona State University Tempe,
More informationI. ACID-BASE NEUTRALIZATION, TITRATION
LABORATORY 3 I. ACID-BASE NEUTRALIZATION, TITRATION Acid-base neutralization is a process in which acid reacts with base to produce water and salt. The driving force of this reaction is formation of a
More informationDisc Diffusion Susceptibility Methods
Disc Diffusion Susceptibility Methods Introduction When a filter paper disc impregnated with a chemical is placed on agar the chemical will diffuse from the disc into the agar. This diffusion will place
More informationDipeptide Synthesis. polarized light (Figure 2).
Dipeptide Synthesis + Scheme 1: Peptide synthesis without carboxyl activation + 2 Throughout your organic chemistry tenure you have been taught the underlying principles necessary to construct simple organic
More informationTamsulosin Hydrochloride Capsules
. nal Revision Bulletin Official October 1, 2011 Tamsulosin 1 standard solution, and shake well. Centrifuge at 1500 rpm for 10 min, and use the supernatant, passing it if Tamsulosin Hydrochloride Capsules
More informationName Lab #3: Solubility of Organic Compounds Objectives: Introduction: soluble insoluble partially soluble miscible immiscible
Lab #3: Solubility of rganic Compounds bjectives: - Understanding the relative solubility of organic compounds in various solvents. - Exploration of the effect of polar groups on a nonpolar hydrocarbon
More informationStructure-Based Design of Covalent Siah Inhibitors
Chemistry & Biology, Volume 20 Supplemental Information Structure-Based Design of Covalent Siah Inhibitors John L. Stebbins, Eugenio Santelli, Yongmei Feng, Surya K. De, Angela Purves, Khatereh Motamedchaboki,
More informationAPPLICATIONS MANUAL. Ifosfamide in blood serum... 2. Ingredients in blood serum... 3. Organophosphorus pesticides in tea leaf... 5
1 APPLICATIONS MANUAL CONTENTS Ifosfamide in blood serum... 2 Ingredients in blood serum... 3 Organophosphorus pesticides in tea leaf... 5 Sudan i ii iii iv in chilli sauce... 6 Pah in water... 7 Phenols
More informationSolid-phase Synthesis of Homodimeric Peptides: Preparation of Covalently-linked Dimers of Amyloid-beta Peptide
Electronic Supplementary Information Solid-phase Synthesis of Homodimeric Peptides: Preparation of Covalently-linked Dimers of Amyloid-beta Peptide W. Mei Kok, a,b,c Denis B. Scanlon, b John A. Karas,
More informationExperiment 8 Synthesis of Aspirin
Experiment 8 Synthesis of Aspirin Aspirin is an effective analgesic (pain reliever), antipyretic (fever reducer) and anti-inflammatory agent and is one of the most widely used non-prescription drugs. The
More informationEnzymes: Amylase Activity in Starch-degrading Soil Isolates
Enzymes: Amylase Activity in Starch-degrading Soil Isolates Introduction This week you will continue our theme of industrial microbiologist by characterizing the enzyme activity we selected for (starch
More informationShort Peptide Synthesis
Short Peptide Synthesis Keith ó Proinsias 8 th February 2010 Introduction Amide bond and basic amide synthesis Solution phase peptide synthesis Protecting groups required for peptide synthesis Coupling
More informationTECHNICAL REPORT STUDY CHEMICAL PROPERTIES OF RATTAN SHOOT FROM PLANTATION IN THAILAND
TECHNICAL REPORT STUDY CHEMICAL PROPERTIES OF RATTAN SHOOT FROM PLANTATION IN THAILAND by Assistant Professor Dr. Noojaree Prasitpan Chemist Analyzer of the ITTO Project on PD 24/00 Rev.1 (I): Promotion
More informationSynthesis of Aspirin and Oil of Wintergreen
Austin Peay State University Department of hemistry hem 1121 autions Purpose Introduction Acetic Anhydride corrosive and a lachrymator all transfers should be done in the vented fume hood Methanol, Ethanol
More informationSupporting Information
Supporting Information Wiley-VCH 2006 69451 Weinheim, Germany Supporting information: experimental details of the synthesis of the amino-functionalized polymers and nanoparticles used Tailor-made ligands
More informationHuman serum albumin (HSA) nanoparticles stabilized with. intermolecular disulfide bonds. Supporting Information
Human serum albumin (HSA) nanoparticles stabilized with intermolecular disulfide bonds Wentan Wang, Yanbin Huang*, Shufang Zhao, Ting Shao and Yi Cheng* Department of Chemical Engineering, Tsinghua University,
More informationLAB 4. Cultivation of Bacteria INTRODUCTION
LAB 4. Cultivation of Bacteria Protocols for use of cultivation of bacteria, use of general growth, enriched, selective and differential media, plate pouring, determination of temperature range for growth
More informationHow To Make A Tri Reagent
TRI Reagent For processing tissues, cells cultured in monolayer or cell pellets Catalog Number T9424 Store at room temperature. TECHNICAL BULLETIN Product Description TRI Reagent is a quick and convenient
More informationQuality. Now Certified to ISO 9001:2008
Quality Now Certified to ISO 90012008 Quality Policy It is Peptides International's goal is to achieve complete customer satisfaction by addressing customer needs and delivering what we promise. The company
More informationMarmara Üniversitesi Fen-Edebiyat Fakültesi Kimya Bölümü / Biyokimya Anabilim Dalı PURIFICATION AND CHARACTERIZATION OF PROTEINS
EXPERIMENT VI PURIFICATION AND CHARACTERIZATION OF PROTEINS I- Protein isolation and dialysis In order to investigate its structure and properties a protein must be obtained in pure form. Since proteins
More informationChapter 5 Classification of Organic Compounds by Solubility
Chapter 5 Classification of Organic Compounds by Solubility Deductions based upon interpretation of simple solubility tests can be extremely useful in organic structure determination. Both solubility and
More informationCarbon. Extraction of Essential Oils from Spices using Steam Distillation. Carbon
Extraction of Essential Oils from Spices using Steam Distillation Contents Objectives 1 Introduction 1 Extraction of trans-anethole from star anise 2 Extraction of trans-cinnamaldehyde from cinnamon 5
More informationExtraction of Caffeine from Energy Drinks
Boston University OpenBU Chemistry http://open.bu.edu Organic Chemistry Laboratory Experiments 2011-07-14 Extraction of Caffeine from Energy Drinks Mulcahy, Seann P. http://hdl.handle.net/2144/1418 Boston
More informationCONFIRMATION OF ZOLPIDEM BY LIQUID CHROMATOGRAPHY MASS SPECTROMETRY
CONFIRMATION OF ZOLPIDEM BY LIQUID CHROMATOGRAPHY MASS SPECTROMETRY 9.1 POLICY This test method may be used to confirm the presence of zolpidem (ZOL), with diazepam-d 5 (DZP-d 5 ) internal standard, in
More informationRecrystallization II 23
Recrystallization II 23 Chem 355 Jasperse RECRYSTALLIZATIN-Week 2 1. Mixed Recrystallization of Acetanilide 2. Mixed Recrystallization of Dibenzylacetone 3. Recrystallization of an Unknown Background Review:
More informationSupporting information. Cyclic peptide-polymer conjugates: grafting to VS grafting from
Supporting information Cyclic peptide-polymer conjugates: grafting to VS grafting from Sophie C. Larnaudie, a Johannes C. Brendel, a,c Katrina A. Jolliffe* b and Sébastien Perrier* a,c a Department of
More informationCULTURE MEDIA FOR COSMETICS INDUSTRY ACCORDING TO ISO STANDARD
CULTURE MEDIA FOR COSMETICS INDUSTRY ACCORDING TO ISO STANDARD PN-EN ISO 11930:2012 Evaluation of the antimicrobial protection of a cosmetic product NEUTRALIZERS 6157 Eugon LT 100 Broth tube 10,9 ml CULTURE
More informationA Simple and efficient method for mild and selective oxidation of propargylic alcohols using TEMPO and calcium hypochlorite
A Simple and efficient method for mild and selective oxidation of propargylic alcohols using TEMPO and calcium hypochlorite Sabbasani Rajasekhara Reddy, a,b Anju Chadha b,c* a School of Advanced Sciences,
More informationSUCRALOSE. White to off-white, practically odourless crystalline powder
SUCRALOSE Prepared at the 41st JECFA (1993), published in FNP 52 Add 2 (1993). Metals and arsenic specifications revised at the 63rd JECFA (2004). An ADI of 0-15 mg/kg bw was established at the 37th JECFA
More informationCH243: Lab 4 Synthesis of Artificial Flavorings by Fischer Esterification
H243: Lab 4 Synthesis of Artificial Flavorings by Fischer Esterification PURPSE: To prepare esters by reaction of carboxylic acids and alcohols. To modify a known procedure to prepare an unknown. DISUSSIN:
More informationEXPERIMENT 9 (Organic Chemistry II) Pahlavan - Cherif Synthesis of Aspirin - Esterification
EXPERIMENT 9 (rganic hemistry II) Pahlavan - herif Materials Hot plate 125-mL Erlenmeyer flask Melting point capillaries Melting point apparatus Büchner funnel 400-mL beaker Stirring rod hemicals Salicylic
More informationSupplementary Information. Primary amino acid lithium salt as a catalyst for asymmetric Michael addition of isobutyraldehyde with β-nitroalkenes.
This journal is (c) The Royal Society of Chemistry 8 Supplementary Information Primary amino acid lithium salt as a catalyst for asymmetric Michael addition of isobutyraldehyde with β-nitroalkenes. Atsushi
More informationTransformation Protocol
To make Glycerol Stocks of Plasmids ** To be done in the hood and use RNase/DNase free tips** 1. In a 10 ml sterile tube add 3 ml autoclaved LB broth and 1.5 ul antibiotic (@ 100 ug/ul) or 3 ul antibiotic
More informationexperiment5 Understanding and applying the concept of limiting reagents. Learning how to perform a vacuum filtration.
81 experiment5 LECTURE AND LAB SKILLS EMPHASIZED Synthesizing an organic substance. Understanding and applying the concept of limiting reagents. Determining percent yield. Learning how to perform a vacuum
More informationoxidize 4-Cholesten-3-one
Isolation of Cholesterol from Egg Yolk Preparation: Bring a hard-boiled egg yolk to lab! Cholesterol (1) is a major component of cell membranes. An egg yolk contains about 200 milligrams of cholesterol,
More informationLab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity
Lab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity Today you analyze the results of your bacterial transformation from last week and determine the efficiency
More information12.1 Hz, 1H), 7.07-7.23 (m, 11H), 7.27-7.33 (m, 6H), 7.54 (d, J = 8.4 Hz, 2H), 10.43 (s, 1H), 13.11 (bs, 1H)
Supporting information Materials and Methods Tetraarylmethane Synthesis THF Br IIIa, b n-buli, Et 2 C Et 2, -78 o C H H Ia, b H CH Ca CH C, 90 o C, 2h aniline.hcl CH CH, toluene, Δ IVa, b IIa, b H 2 =
More informationEXPERIMENT 5: DIPEPTIDE RESEARCH PROJECT
EXPERIMENT 5: DIPEPTIDE RESEARCH PROJECT Pre-Lab Questions: None. 64 I. Background Information DIPEPTIDE RESEARCH PROJECT Methods developed by organic chemists for the synthesis of biopolymers have had
More informationSpatial Screening of Cyclic Neoglycopeptides: Identification of Multivalent Wheat Germ Agglutinin Ligands**
1 Spatial Screening of Cyclic Neoglycopeptides: Identification of Multivalent Wheat Germ Agglutinin Ligands** Valentin Wittmann* and Sonja Seeberger Experimental Section General. Solid-phase peptide synthesis
More informationSeparation by Solvent Extraction
Experiment 3 Separation by Solvent Extraction Objectives To separate a mixture consisting of a carboxylic acid and a neutral compound by using solvent extraction techniques. Introduction Frequently, organic
More informationRunning protein gels and detection of proteins
Running protein gels and detection of proteins 1. Protein concentration determination using the BIO RAD reagent This assay uses a colour change reaction to give a direct measurement of protein concentration.
More informationTECHNICAL BULLETIN. HIS-Select Nickel Affinity Gel. Catalog Number P6611 Storage Temperature 2 8 C
HIS-Select Nickel Affinity Gel Catalog Number P6611 Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description HIS-Select Nickel Affinity Gel is an immobilized metalion affinity chromatography (IMAC)
More informationDetermining the Quantity of Iron in a Vitamin Tablet. Evaluation copy
Determining the Quantity of Iron in a Vitamin Tablet Computer 34 As biochemical research becomes more sophisticated, we are learning more about the role of metallic elements in the human body. For example,
More informationLight-driven Nanoscale Chiral Molecular Switch: Reversible Dynamic Full Range Color Phototuning
This journal is (c) The Royal Society of Chemistry Supporting Information Light-driven Nanoscale Chiral Molecular Switch: Reversible Dynamic Full Range Color Phototuning Ji Ma, Yannian Li, Timothy White,
More informationHiPer Ion Exchange Chromatography Teaching Kit
HiPer Ion Exchange Chromatography Teaching Kit Product Code: HTC001 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 5-6 hours Storage Instructions: The kit is stable for
More informationHEMOLYTIC PROPERTY AND GC MS ANALYSIS OF COCOS NUCIFERA FIBER EXTRACTS FROM MARINE COASTAL AREA
Innovare Academic Sciences International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 6, Issue 4, 2014 Original Article HEMOLYTIC PROPERTY AND GC MS ANALYSIS OF COCOS NUCIFERA FIBER
More informationInduction of Enzyme Activity in Bacteria:The Lac Operon. Preparation for Laboratory: Web Tutorial - Lac Operon - submit questions
Induction of Enzyme Activity in Bacteria:The Lac Operon Preparation for Laboratory: Web Tutorial - Lac Operon - submit questions I. Background: For the last week you explored the functioning of the enzyme
More informationWorking with Hazardous Chemicals
A Publication of Reliable Methods for the Preparation of Organic Compounds Working with Hazardous Chemicals The procedures in Organic Syntheses are intended for use only by persons with proper training
More informationCovalent Conjugation to Cytodiagnostics Carboxylated Gold Nanoparticles Tech Note #105
Covalent Conjugation to Cytodiagnostics Carboxylated Gold Nanoparticles Tech Note #105 Background Gold nanoparticle conjugates have been widely used in biological research and biosensing applications.
More informationTechnical Report. Automatic Identification and Semi-quantitative Analysis of Psychotropic Drugs in Serum Using GC/MS Forensic Toxicological Database
C146-E175A Technical Report Automatic Identification and Semi-quantitative Analysis of Psychotropic Drugs in Serum Using GC/MS Forensic Toxicological Database Hitoshi Tsuchihashi 1 Abstract: A sample consisting
More informationInternational Journal of Research and Reviews in Pharmacy and Applied science. www.ijrrpas.com
International Journal of Research and Reviews in Pharmacy and Applied science www.ijrrpas.com P.V.V.Satyanarayana*, Alavala Siva Madhavi NEW SPECTROPHOTOMETRIC METHODS FOR THE QUANTITATIVE ESTIMATION OF
More informationOrganic Chemistry Lab Experiment 4 Preparation and Properties of Soap
Organic Chemistry Lab Experiment 4 Preparation and Properties of Soap Introduction A soap is the sodium or potassium salt of a long-chain fatty acid. The fatty acid usually contains 12 to 18 carbon atoms.
More informationCypExpress 3A4 Catalyzed Conversion of Testosterone (TE) to 6β- Hydroxytestosterone (HT)
TM CASE STUDY CypExpress 3A4 Catalyzed Conversion of to Shuvendu Das, 1 Enrique Martez, 2 and Mani Subramanian 1 1 Center for Biocatalysis and Bioprocessg, University of Iowa 2 Oxford Biomedical Research,
More informationLUMEFANTRINE Draft proposal for The International Pharmacopoeia (October 2006)
October 2006 RESTRICTED LUMEFANTRINE Draft proposal for The International Pharmacopoeia (October 2006) DRAFT FOR DISCUSSION World Health Organization 2006 All rights reserved. This draft is intended for
More informationGuide to Reverse Phase SpinColumns Chromatography for Sample Prep
Guide to Reverse Phase SpinColumns Chromatography for Sample Prep www.harvardapparatus.com Contents Introduction...2-3 Modes of Separation...4-6 Spin Column Efficiency...7-8 Fast Protein Analysis...9 Specifications...10
More informationSelf-Propelled Chemotactic Ionic Liquid Droplets
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2014 Supporting Information Self-Propelled Chemotactic Ionic Liquid Droplets Wayne Francis, Cormac Fay,
More informationCLINICAL STUDY OF MARKETED AYURVEDIC PREPARATION IN THYROID INDUCED STRESS A. A. ROY* 1, K. R. KAKADE 2, R.P. BHOLE 1, V. B.
International Journal of PharmTech Research CODEN( USA): IJPRIF ISSN : 0974-4304 Vol.1, No.3, pp 870-874, July-Sept 2009 CLINICAL STUDY OF MARKETED AYURVEDIC PREPARATION IN THYROID INDUCED STRESS A. A.
More informationTable of Content. Enzymes and Their Functions Teacher Version 1
Enzymes and Their Functions Jeisa Pelet, Cornell University Carolyn Wilczynski, Binghamton High School Cornell Learning Initiative in Medicine and Bioengineering (CLIMB) Table of Content Title Page Abstract..
More informationEnantiomers: Synthesis, characterization, and resolution of tris(ethylenediamine)cobalt(iii) chloride Introduction:
Enantiomers: Synthesis, characterization, and resolution of tris(ethylenediamine)cobalt(iii) chloride Introduction: The development of coordination chemistry prior to 1950 involved the synthesis and characterization
More informationIsolation of Caffeine from Tea
Isolation of Caffeine from Tea Introduction A number of interesting, biologically active compounds have been isolated from plants. Isolating some of these natural products, as they are called, can require
More informationSupplemental data. A simple and effective cleavable linker for chemical proteomics applications
Supplemental data A simple and effective cleavable linker for chemical proteomics applications Yinliang Yang, annes ahne, Bernhard Kuster, and Steven. L. Verhelst * Figure S1 Figure S2 Figure S3 Table
More informationTECHNICAL BULLETIN. FluoroTag FITC Conjugation Kit. Product Number FITC1 Storage Temperature 2 8 C
FluoroTag FITC Conjugation Kit Product Number FITC1 Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description The FluoroTag FITC Conjugation Kit is suitable for the conjugation of polyclonal and
More informationHands-On Labs SM-1 Lab Manual
EXPERIMENT 4: Separation of a Mixture of Solids Read the entire experiment and organize time, materials, and work space before beginning. Remember to review the safety sections and wear goggles when appropriate.
More informationPlant Genomic DNA Extraction using CTAB
Plant Genomic DNA Extraction using CTAB Introduction The search for a more efficient means of extracting DNA of both higher quality and yield has lead to the development of a variety of protocols, however
More informationStandard practices for Fmoc-based solid-phase. peptide synthesis in the Nowick laboratory. (Version 1.6.1)
Standard practices for Fmoc-based solid-phase peptide synthesis in the Nowick laboratory (Version 1.6.1) Adam G. Kreutzer and Patrick J. Salveson E-mail: Contents Contributions to this guide 3 General
More informationFighting the Battles: Conducting a Clinical Assay
Fighting the Battles: Conducting a Clinical Assay 6 Vocabulary: In Vitro: studies in biology that are conducted using components of an organism that have been isolated from their usual biological surroundings
More informationQuantifying Bacterial Concentration using a Calibrated Growth Curve
BTEC 4200 Lab 2. Quantifying Bacterial Concentration using a Calibrated Growth Curve Background and References Bacterial concentration can be measured by several methods, all of which you have studied
More informationHigh Performance Thin Layer Chromatographic Method for Estimation of Cefprozil in Tablet Dosage Form
ISSN: 0973-4945; CODEN ECJHAO E- Chemistry http://www.e-journals.net Vol. 5, No.3, pp. 427-430, July 2008 High Performance Thin Layer Chromatographic Method for Estimation of Cefprozil in Tablet Dosage
More information